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Originally published In Press as doi:10.1074/jbc.M100493200 on May 7, 2001

J. Biol. Chem., Vol. 276, Issue 28, 25910-25918, July 13, 2001
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Terminal Minihelix, a Novel RNA Motif That Directs Polymerase III Transcripts to the Cell Cytoplasm
TERMINAL MINIHELIX AND RNA EXPORT*

Carole GwizdekDagger §, Edouard Bertrand||**, Catherine Dargemont§, Jean-Claude LefebvreDagger , Jean-Marie Blanchard||, Robert H. Singer**, and Alain DoglioDagger Dagger Dagger

From the Dagger  U526-Laboratoire de Virologie, Faculté de Médecine, Avenue de Valombrose, 06107 Nice cedex 2, France, the || Institut de Génétique Moléculaire de Montpellier, 34293 Montpellier Cedex 5, France, the ** Department of Anatomy and Structural Biology and Cell Biology, Albert Einstein College of Medicine, Bronx, New York 10461, and the § Laboratoire de Transport Nucléocytoplasmique, Institut Jacques Monod, 2 place Jussieu, 75251 Paris cedex 5, France

Determining the cis-acting elements controlling nuclear export of RNA is critical, because they specify which RNA will be selected for transport. We have characterized the nuclear export motif of the adenoviral VA1 RNA, a small cytoplasmic RNA transcribed by RNA polymerase III. Using a large panel of VA1 mutants in both transfected COS cells and injected Xenopus oocytes, we showed that the terminal stem of VA1 is necessary and sufficient for its export. Surprisingly, we found that the nucleotide sequence within the terminal stem is not important. Rather, the salient features of this motif are its length and its relative position within the RNA. Such stems thus define a novel and degenerate cytoplasmic localization motif that we termed the minihelix. This motif is found in a variety of polymerase III transcripts, and cross-competition analysis in Xenopus oocytes revealed that export of one such RNA, like hY1 RNA, is specifically competed by VA1 or artificial minihelix. Taken together these results show that the minihelix defines a new cis-acting export element and that this motif could be exported via a novel and specific nuclear export pathway.

* This work was supported by the Agence Nationale de Recherche sur le SIDA (ANRS), Sidaction, the Boris Vlasov Fundation (Monaco), ARC grant 9043, and National Institutes of Health Grants GM54887 and 57071.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Both authors contributed equally to this work.

Dagger Dagger To whom correspondence should be addressed. Tel.: 33 4-93-37-7678; Fax: 33 4-93-81-5484; E-mail: doglio@unice.fr.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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